Most commercial therapeutic proteins, such as monoclonal antibodies (Mabs), are produced from traditional hybridoma cultures or genetically engineered Chinese hamster ovary (CHO) cells using large scale : bioreactors. Bioprocessing of therapeutic proteins is exceedingly expensive because of the high costs of , building and maintaining FDA-approved facilities, complex serum-free media, and talented labor. There is a tremendous market need for methods that can enhance protein production that ideally complement the current methods of production. The overall goal of the proposed project is to evaluate the effectiveness of senescence-enhanced protein production (RP Shift) in commercially relevant bioreactor setting. In the Phase I portion .of this project, Mab production was enhanced nearly 20-fold from RP Shift-competent hybridoma cells in small flasks. The Phase II portion of this project addresses the effectiveness of the RP Shift in commercially relevant bioreactors, the most commonly used being batch and flow-through systems. The aims of this Phase II proposal are to monitor commercially relevant endpoints from RP Shift-competent CH450 and MH70 hybridoma cells that were engineered in the Phase I portion of the project. Specifically, 1) enhanced productivity of Mab (in pg/cell/day monitored daily), 2) increased culture lifetime measured in days), and 3) the stability and integrity of the Mab produced will be measured. First cell lines with increased productivity of at least 2-fold during RP Shift in small volume test systems will be isolated. Of these RP Shift competent cell lines, the best two will be transferred to commercial scale bioreactors and examined for enhanced protein production and increased bioreactor lifetimes. It is expected that enhancements of at least 2-fold in large-scale bioreactors will attract interest from biopharmaceutical manufacturers. During Phase III, the RP Shift technology and ready-to-go RP Shift-competent cell lines will be licensed to biopharmaceutical manufacturers in an effort to reduce costs for them and ultimately the general public. [unreadable] [unreadable]